Effect of pumpkin seed Intake Cucurbita l. maximum on the lipid profile and Glucose in male Wistar rats [1]

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ROSBACH, Tiago [2], PEREIRA, Francine Martins [3]

ROSBACH, Tiago; PEREIRA, Francine Martins. Effect of pumpkin seed Intake Cucurbita l. maximum on the lipid profile and Glucose in male Wistar rats. Multidisciplinary Core scientific journal of knowledge. 03 year, Ed. 06, vol. 05, pp. 5-16, June 2018. ISSN:2448-0959

Summary

The phytosterol and cholesterol are present in foods of animal and plant origin respectively being cholesterol responsible for cellular permeability acts in the synthesis of bile acids, vitamin D and hormones. In the human body about of 70% of the endogenous cholesterol, and only 30% is of exogenous origin, acquired through the diet. The present work aims to evaluate the effects of pumpkin seed ingestion on glucose and lipid metabolism of male Wistar rats. The used-if 30 Wistar rats with hypercholesterolemia induction for 20 days. Blood was collected for proper examination, where we used the non-enzymatic colorimetric test kit for cholesterol, TGS, LDL, VLDL, HDL and blood glucose. For the following work was compared with the hipercolesterolêmico group hipercolesterolêmico group more pumpkin seed using the t test for nonparametric samples with significance value of 0.05, where < observed increase in the HDL fraction cholesterol, but for the other variables not obtained significant results. With the study, it was possible to conclude that the pumpkin seed because it is high in fiber and fitoesterois when added to food of Wistar rats, was able to increase the HDL cholesterol.

Keywords: Cucurbita maximum L, Wistar rats, Hypercholesterolemia.

Introduction

Phytotherapy and the use of plants for medicinal purposes are part of the practice of folk medicine, being a set of knowledge and beliefs passed from person to person especially by oral tradition. Due to the industrialization process this ancient practice comes down.

However the use of plants is being touted as an effective primary health care, and may complement the treatment conventionally employed, for the lower income population (ELDIN S, et al. 2001).

The investigation of the use of plants by the indigenous communities for the purpose of treatment made possible the discovery of many drugs with psychoactive action, like yohimbine, ephedrine, tubocurarine and galantamina. In this context the use of plants for medicinal purposes is the field of Ethnopharmacology that combines knowledge of popular/traditional use with chemical and pharmacological studies as a discovery of new drugs (ELISABETSKY, 2007).

Natural products demonstrate a great significance in the discovery of new biologically active compounds and the development of new medicines for the treatment of numerous diseases. In the year 2000, the ratio of drugs developed from natural products was around 20%, with a significant increase in the following years. Newman (2008) suggested that about 60% of the drugs available were derived from natural products, being that they are an important source for the development of new agents with potential. In addition, it has been shown that many substances derived from nature, can interact with mammalian receptors attesting the bioactivity of these products.

Cholesterol is a type of fat (lipid) found naturally in our body, fundamental to your normal operation. Cholesterol is the structural component of cell membranes throughout the body and is present in the brain, nerves, muscles, skin, liver, intestines and heart. Our body uses cholesterol to produce many hormones, vitamin D and bile acids that help in the digestion of fats. 70% of the endogenous cholesterol being produced by our own organism, in the liver, with only 30% of exogenous origin through diet (BRAGAGNOLO, 2001).

The hipercolesterolêmia occurs mainly by ingestion of saturated fatty acid myristic found in fats like animal found in coconut oil lauric and Palmitic in olive oil-palm oil. Unsaturated fatty acids are divided into two groups, polyunsaturated (Omega-3 and Omega-6), where your beneficial effect is to decrease the number of LDL receptors, thus reducing the risk of cardiovascular disease and monounsaturated (Omega 9) which decreases the total cholesterol and LDL cholesterol, in addition to possessing antithrombotic effect and inhibit platelet aggregation (CUPPARI, 2002).

Dyslipidemias are the lipid metabolic changes resulting from disturbances in any phase of lipid metabolism that cause impact on serum levels of lipoproteins. The dyslipidemias are among the most important risk factors for atherosclerotic cardiovascular disease, along with hypertension, obesity and diabetes mellitus. However, national and international studies show reduction in coronary events when is intervention in risk factors (CASTELLI WP, 1984).

The total cholesterol and LDL cholesterol are directly related to morbidity and mortality from coronary artery disease. The isolated hypertriglyceridemia does not constitute independent risk factor of coronary heart disease, but happens to have him when associated with high levels of LDL-c and low level of HDL-c. Have HDL-c (higher-density lipoprotein cholesterol), operates in reverse cholesterol transport, reducing or stopping the formation of plaque. At the third National Health and Nutrition Examination (NHANES III), held in the United States, the factors affecting serum cholesterol levels were numerous, including the age, diet, genetic predisposition and the body weight (STEINBERG D .1987).

The dietary fiber is the edible part of plant or carbohydrates that are resistant to digestion and absorption in the small intestine of humans, with complete or partial fermentation in the large intestine. The dietary fiber includes polysaccharides, cellulose, hemicelluloses as vegetables, pectins, gums and mucilage, oligosaccharides, lignin and associated plants substances. This definition was drawn up in 1999 by the American Association of Cereal Chemists, after many debates subsidized with information from industries, academia and Government agencies of various countries (COPPINI, l. Z, et al. 2002).

Dietary fibres are divided into two categories because of solubility in water, they are soluble fiber and insoluble fiber. The soluble fibre include most pectin, gums, mucilage and hemicelluloses. Are found in fruits, oat bran, barley and legumes (beans, lentils, peas and chickpeas) (COPPINI LZ, et al. 2002, CHANDRA MLF .1997).

Soluble fiber has as a characteristic to increase the intestinal transit time and is related to the decrease of gastric emptying, delay the absorption of glucose, postprandial blood glucose reduction and reduction of blood cholesterol due to their absorption properties checking content viscosity luminal. (COSTA RP, et al. 1997).

Insoluble fibers contributing to the increase in the volume of fecal cake, reduced intestinal transit time, delay absorption of glucose and starch hydrolysis retardation. Cellulose, lignin, some hemicelluloses and mucilage are examples of insoluble fibers. Are found in greater quantity in the fruit peels in the wheat bran in whole grains and their products, in the roots and in the vegetables. Generally, do not undergo fermentation, so that, when this occurs, it occurs slowly. Proportionally, the insoluble fraction of the fibers is the most abundant, constituting about of 2/3 the 3/4 of dietary fiber to a diet consisting of varied plant foods (MARLETT JA, SLAVIN JL .1997).

Phytosterols as well as cholesterol are constituents of cell membranes in the case of plants and are responsible for the permeability of the cell. Structurally differs from cholesterol only because it contains one or two methyl or ethyl groups on the molécula1 side chain. Only recently the phytosterols aroused the interest of researchers due to your potential benefit in reducing serum cholesterol levels, its effects as natural part of common foods are moderate, since your current consumption is low (between 200 and 400 mg/day). But your addition to foods have proven results in reducing plasma cholesterol (CHAN Y, v. et al. 2006).

The phytosterols are active at various levels in cholesterol metabolism, although your mechanism of action is not yet fully elucidated (SANÉ, et al. 2006).

In the intestinal lumen phytosterol and cholesterol are incorporated into micelles entering the enterocytes by brush surround the transporter NPC1L1. So is cholesterol esterified by acyl CoA: cholesterol acyltransferase (ACAT) and incorporated into chylomicrons, which are secreted to the lymph. This happens to a small part of the fitoesterois as they have little affinity for the ACAT. That is, while the cplesterol is absorbed, the fitoesterois are very little. Only 0.5 to 1.9% of phytosterols are absorbed. Once in the enterocyte a small part of the cholesterol and the large proportion of phytosterols are excreted again into the intestinal lumen ABCG5 and ABCG8 transporters through so do not accumulate in the body (YANG C, et al. 2004).

Among the varied sources of fiber-rich foods include pumpkin seed being a by-product of pumpkin. The pumpkin with the order Cucurbitales, family Cucurbitaceae and Cucurbita species, being the state your mature customarily used to compose the diet. The pumpkin seed has also been used in brazilian folk medicine. Studies in animals show that your fresh consumption, without suffering heat treatment, can reduce the bioavailability of some nutrients due to presence of phytates. (DEL-VECCHIO, et al 2005).

The pumpkin seed has been used in many ways in human consumption salt and toast as an appetizer, oil or flour as coadjuvant in the thinning due to your high level of dietary fibre, it also has anthelmintic effect and antioxidant, In addition to being a good protein source. (AL-ZUHAIR H, FATTAH AAA, SAYED MI .2000).

Due to the positive contribution of dietary fiber to health and to the beneficial effects of the fitoesterois and the high fiber content of pumpkin seed the aim of this study was to evaluate the effects of pumpkin seed on lipid and glucose metabolism in Wistar rats males.

Methodology

The experiment began with the induction to hypercholesterolemia and subsequently the offer of pumpkin seed, the animals were sacrificed at the end of the experiment. Altogether 30 male Wistar rats were used in approximately 90 days of age from the vivarium of Faculdade Assis Gurgacz. In the course of the experiment the animals were kept in 6 boxes with shavings containing 5 animals in each, cleaned 3 times a week for 7 weeks and weighed weekly. Were divided into 3 groups: control group, hipercolesterolêmico group and hipercolesterolêmico group more pumpkin seed.

For the induction of hypercholesterolemia was used mixed ration in swine fat, with the proportion of 200 g/50 g fat ration for 15 ', during 30 days. After this period the groups induced hypercholesterolemia received along with the feed and bathes in the proportion of 1 kg of feed/100 g lard over the pumpkin seed pumpkin seed group.

Pumpkin seeds were acquired in natural products stores and offered 3 times a week, where each animal received 0, 2 g according to the instructions described on the label of the product and divided by the average weight of rats during 25 days.

For the collection of the blood sacrifice of animals through a guillotine to beheading, which are caught one by one, the blood was deposited in collection tube with K ³ EDTA (anticoagulant solution for use in hematology) silicone not with the help of a funnel and taken the centrifuge for 15 ´ RBC plasma separation, then the plasma was collected with the help of a syringe and transferred to a microcentrifuge tube.

To evaluate the results it was verified the body weight, HDL, LDL, VLDL, total cholesterol and triglycerides by analyzing changes in plasma levels occurred through biochemical tests and the total weight.

To check the variables, total cholesterol, triglycerides, LDL, VLDL and HDL cholesterol was used and colorimetric enzymatic test kits for determination in serum and/or plasma Bioliquid brand of the company Laborclin, by following the procedures described in the package insert and calculations (absorbance of sample/absorbance standard x 200) for total cholesterol and triglycerides and HDL cholesterol (absorbance of supernatant/standard x 50 x 1.1 absorbance) where the liquid is removed from the supernatant plasma after centrifugation to separate the fat. For LDL cholesterol (HDL LDL + VLDL +) and VLDL cholesterol (TG/5) where TG is triglyceride form. The absorbance values were given by espectofotômetro with wavelength of 500nm.

For the following work Was compared the pumpkin seed group with the hipercolesterolêmico group using the t test for nonparametric samples with significance value of p < 0.05.

Results and discussion

Through the results showed a significant increase in HDL cholesterol fraction in hipercolesterolêmico group more pumpkin seed in relation to that group only received hypercholesterolemic diet, for the variables total cholesterol, triglycerides, LDL, VLDL and glucose there were no significant results as shown in Figure 1.

Figure 2: effect of pumpkin seed Administration on the total body weight of Wistar rats hypercholesterolemic. p < 0.0001.
Figure 2: effect of pumpkin seed Administration on the total body weight of Wistar rats hypercholesterolemic. p < 0.0001.
Figure 1: effect of pumpkin seed administration in hypercholesterolemic male Wistar rats. The values for the variables total cholesterol, HDL, LDL, VLDL, TGS and glucose are expressed in mg/dl. * p < 0.05. HDL: high density lipoprotein, LDL: low density lipoprotein; VLDL: intermediate density lipoprotein; TGS: triglycerides.
Figure 1: effect of pumpkin seed administration in hypercholesterolemic male Wistar rats. The values for the variables total cholesterol, HDL, LDL, VLDL, TGS and glucose are expressed in mg/dl. * p < 0.05. HDL: high density lipoprotein, LDL: low density lipoprotein; VLDL: intermediate density lipoprotein; TGS: triglycerides.

As demonstrated in Figure 1 the values of total cholesterol, LDL cholesterol, VLDL, triglycerides and glucose were not significant, but in the study of Borges et al. as for the 2012 triglycerides diets with flaxseed generated ducker effect, when compared to the control group (p < 0.005), it was noted in the VLDL fraction. You can analyze even in a high-fat diet, supplementation with flaxseed presented significant beneficial effect on LDL cholesterol fraction, with lower values, when compared with the control group.

In a study conducted in the year 1998 by Sgarbieri during 28 days & Cold tested the effect of diets containing 0%, 10% and 15% of guar gum on the lipid profile of wistar rats where observed that at concentrations equal to or greater than 10% guar gum was effective in the treatment of hypercholesterolemia in mice.

In work done by Derivi and Pourchet-fields (1984), studying two varieties of beans: Carpenter bean (Phaseolus vulgaris) and macassar bean (Vigna sinensis), pectin has been identified as a hipocolesterolêmico component present in legumes. The authors found that the sharp hipocolesterolêmico presented by carioca bean effect was due to the presence of a high content of soluble (pectin 1.46 g%), whereas this effect was not observed in macassar bean which featured 0.64 g% pectin soluble in your composition.

In study by Fietz & Salgado (1999), by adding the diet of Wistar rats pectin and cellulose by 30 days containing 5, 10, 15 and 20% of these, observed that diets with more than 10% of pectin and cellulose had significant effect in reducing levels of cholesterol.

In a human study, there was significant reduction of plasma total cholesterol levels and LDL cholesterol from consumption, for a period

corresponding to 21 days, the esters of sterols (12.9 and 13.4%, respectively) compared with esters of stanols (10.2 and 7.9%, respectively). The control group was submitted to the consumption of margarine without addition of phytosterols (6.0 and 3.9%, respectively) (P < 0.05). Plasma levels of HDL cholesterol and triglycerides, were not observed significant changes due to the use of phytosterols (CLIFTON, 2002).

In study by Cerqueira et al., (2008), it was observed that after ingestion of pumpkin seed meal there was a reduction in blood sugar levels compared to control group (p < 0.05).

There was a significant increase in relation to the body weight of the mice that received the most hypercholesterolemic diet pumpkin seed compared to mice with hypercholesterolemic diet as shown in Figure 2.

Figure 2: effect of pumpkin seed Administration on the total body weight of Wistar rats hypercholesterolemic. p < 0.0001.
Figure 2: effect of pumpkin seed Administration on the total body weight of Wistar rats hypercholesterolemic. p < 0.0001.

The results can be observed that the rats fed on hypercholesterolemic diet more pumpkin seed had significant increase in total body weight (*** p < 0.0001) in relation to the hipercolesterolêmico group, this is due to the fact that the fats have the power to give more flavor to foods making them tastier and the juicy sweetness of pumpkin seed making the tastiest ration increasing consumption by rats.

Second Akiyama et al (1996), mice with hiperenergética and diet with high content of fats tend to develop obesity. In study Duarte et al. (2006) reported that, even without increasing the daily amount of food ingested, the high-fat diet promoted obesity.

To analyze the total energy consumption over the next eight weeks, higher energy intake was observed in the experimental group the enough for the accumulation of adipose tissue, as noted in the work of Pullar & Webster (1977).

The literature indicates that not only the totals of energy ingested and regulate the amount of stocks. The balance of each macronutrient seems to possess a rigorous control to adjust your intake with your oxidation (and vice versa) and maintain a State of balance. The balance of nitrogen and carbohydrates is facilitated by the ability of the body to adjust the rates of oxidation of glucose and amino acids, respectively, in relation to its food consumption. In the case of fats, this adjustment is much less precise and the increase in consumption does not stimulate your the your oxidation, and the increase in lipid intake will induce lipid balance sheet positive and, therefore, to build up in body fat mass (PEREIRA et al., 2003).

Conclusion

On the basis of the obtained results it can be concluded that the pumpkin seed can be a good ally in controlling hypercholesterolemia due to your composition rich in fiber and fitoesterois that has proven beneficial action in other studies. The increase in HDL cholesterol fraction is because of the presence of linoleic acid (Omega 6). The other variables may not present significance due to the fact that the quantity supplied or that there was no control of how much each rat ate.

For future works we suggest increasing the dose of pumpkin seed offered and also increase the number of days you receive the product by passing to offer every day of the week.

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[1] Final project presented as a requirement for obtaining a Bachelor's degree in nutrition.

[2] Academic-Faculdade Assis Gurgacz.

[3] Faculty adviser-Faculdade Assis Gurgacz.

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